Overview and Rationale for use
Fat Smack™ is a new product designed by PURUS LABS®, which adheres to the established guidelines of PURUS LABS® in terms of including those ingredients that have been used in humansubjects via oralingestion, and reported to promote favorable outcomes in relation to the target areas of interest. Fat Smack™ contains a unique blend of ingredients, each carefully selected for their potential impact on weight/fat loss related variables, used at the correct dosages based on the scientific studies reviewed (again, using human subjects).

The following is the firm position of PURUS LABS® in formulating dietary supplements: If only a few ingredients can be identified as potentially efficacious in terms of the desired outcome measures (which is indeed the case when reviewing the scientific literature), then it’s best to simply include only those potent few ingredients at the correct dosages as opposed to adding tons of “window dressing” for purposes of marketability. In short, the plan adopted by PURUS LABS® is simple—brevity and precision. The text below outlines the ingredients used within Fat Smack™, while providing a brief summary of the rationale for use.

1,3,7-trimethylxanthine (Caffeine)
Commonly referred to as caffeine, 1,3,7-trimethylxanthine is very well-studied and extensively used. In fact, caffeine is the most widely consumed pharmacological substance in the world. In the United States, caffeine consumption averages ~200 mg daily, which is the equivalent of approximately two cups of coffee. Of course, individuals consuming more concentrated beverages (e.g., espresso, energy drinks, etc.) are likely ingesting amounts much higher than this.

The most well-described physiological effects of caffeine relate to enhanced mood, cognition, exercise performance, and weight loss associated outcomes (e.g., increased thermogenesis [Belza et al., 2009], increased free fatty acid release and catecholamine levels [Patwardhan et al., 1980], decreased appetite). Multiple mechanisms are associated with the ergogenic effect of caffeine, including improved cognitive function, increased catecholamine secretion and associated lipolysis, enhanced Na+/K+ pump activity which serves to enhance excitation contraction coupling, increased calcium mobilization, phosphodiesterase inhibition, and antagonism of adenosine receptors. In the same way, many of these effects may be responsible for the potential weight/fat loss effects of caffeine, a complex area of investigation that has previously been reviewed elsewhere (Acheson et al., 2004). While individual response to caffeine varies based on tolerance, dosages in the literature have generally ranged from 3-6 mg∙kg-1 body mass, with individuals who do not frequently use caffeine generally responding best (Bell and McLellan, 2002). With this understanding, it is possible that “cycling” on and off caffeine (as well as other stimulants) may provide for the best effects (as opposed to regular intake without time spent away from treatment), in terms of both efficacy and safety. In general, caffeine is well-tolerated by most individuals, approved for use as a food ingredient, and is considered safe with moderate consumption (Heckman et al., 2010).

2-amino-4-methylhexane
The ingredient 2-amino-4-methylhexane is a component of geranium oil (Ping et al., 1996), also referred to as 1,3-Dimethylamylamine, 1,3-Dimethylpentylamine, Methylhexaneamine, 4-Methyl-2-hexylamine, and most commonly, in the lay press, as “geranamine”. The name geranamine has been trademarked by Proviant Technologies, Inc. in 2005 (US trademark number: 78542697) and indicated as a “Constituent of flower oil sold as an integral component of nutritional supplements.” 2-amino-4-methylhexane appears to provide a sympathomimetic effect in human subjects; that is, it is thought to mimic the effects of the sympathetic nervous system such as the chemicals epinephrine, norepinephrine, and dopamine. This does not indicate that it necessarily elevates these chemicals; rather, it may mimic their effects. These may include enhanced fatty acid mobilization from storage depots, as well as providing a feeling of euphoria. It should be noted that very little is known about this ingredient, despite its widespread use within the sport supplement market. In fact, this is the one ingredient contained within Fat Smack™ that is not yet supported by peer-reviewed, published clinical data. However, research pertaining to its use in human subjects is ongoing. In the meantime, anecdotal reports exist for improved focus, mood, exercise performance, and appetite suppression.

In preliminary work with this agent, 2-amino-4-methylhexane combined with caffeine has been reported to provide positive effects on resistance exercise performance in men, as well as reduced appetite in both men and women (unpublished findings). That being said, it should be noted that the combination of caffeine (250mg) and 2-amino-4-methylhexane (50mg), when ingested by healthy men and women, also results in an average blood pressure increase of 10-14mmHg (unpublished findings), similar to that observed with higher dosages of caffeine alone or with caffeine combined with other stimulants (e.g., ephedra). Such an increase in blood pressure is likely the case when using many of the over-the-counter weight/fat loss and stimulant-containing performance supplements; regardless, PURUS LABS® understands the importance of full disclosure of facts once they become available. It is imperative that individuals considering the use of Fat Smack™, or any other products containing a combination of caffeine and 2-amino-4-methylhexane, understand their blood pressure may acutely increase with use. Based on this fact, those individuals with elevated blood pressure (>120/80mmHg) should not consider using this product or any other products that contain stimulants. While blood pressure appears to be elevated acutely with a single dosage of 2-amino-4-methylhexane, it is unknown whether or not resting blood pressure will be elevated following chronic use (i.e., daily); more research is needed to determine such effects. Finally, because both caffeine and 2-amino-4-methylhexane are classified as stimulants, it is possible that individuals using these agents (in particular 2-amino-4-methylhexane—which is now listed on the banned substance list for many sports organizations) may increase the likelihood of testing positive for a banned substance in sports competition or other arenas. It should be noted that the amount and duration of both caffeine and 2-amino-4-methylhexane intake necessary to lead to a “positive” test is presently unknown.

The dosage of 2-amino-4-methylhexane contained within Fat Smack™ is based on observations in human subjects using 2-amino-4-methylhexane alone and in combination with caffeine, as well as pilot testing done by PURUS LABS® in a variety of healthy men and women with normal blood pressure. While the combined dosage of these ingredients contained within one capsule of Fat Smack™ is adequate for many individuals, some may opt to use the full serving of two capsules per day (together or in divided dosages). Anecdotally, it does appear the overall “stimulant” and blood pressure raising effects of 2-amino-4-methylhexane are somewhat attenuated with continued used; however, this requires laboratory-based study. Further research is indeed warranted in relation to 2-amino-4-methylhexane and its role in providing favorable effects related to mood, fatty acid mobilization, exercise performance, and weight/fat loss over time. Moreover, additional safety data are necessary related to this agent.

Epigallocatechin Gallate (EGCG)
Epigallocatechin gallate (EGCG), sometimes referred to as epigallocatechin 3-gallate, is one of several catechins, and is the ester of epigallocatechin and gallic acid. EGCG is found in relatively high amounts within tea (green tea in particular), has multiple physiological properties ranging from antioxidant activity to fueling weight/fat loss (Saito et al., 2009), and is generally viewed as a candidate ingredient in the treatment of metabolic syndrome (Thielecke and Boschman, 2009). Considerable interest has been given to green tea extract (and specifically the catechins within the green tea—EGCG in particular) as a cardio-protective agent with significant potential to minimize oxidative stress—a condition which has been associated with multiple known human diseases as well as the aging process (Giustarini et al., 2009). In short, the beneficial effects of green tea consumption have been reported in multiple peer reviewed manuscripts.

It should be understood that despite the fact that EGCG is found within tea, the use of isolated amounts of EGCG is not the same as simply drinking a cup of brewed or ready-to-drink green tea. The amount of EGCG contained within tea can vary considerably depending on the exact type of tea, whether it is caffeinated or decaffeinated, whether it is flavored or not, whether it is brewed or ready-to-drink, etc. (USDA report). Related to the above, natural brewed green tea appears best, with an average EGCG content of ~78mg per 100g of edible portion (USDA report). To obtain the research supported dosage of EGCG (~300mg/day), it would be wise to regularly consume natural brewed green tea or to consume an EGCG encapsulated product to ensure the correct dosage is provided.

In terms of biological activity and absorption of EGCG when provided as a green tea extract versus actual tea consumption, the following should be noted: First, in a study in which green tea extract was compared with black and green tea (providing the same amount of EGCG), flavanol absorption was enhanced when tea polyphenols were administered as a green tea supplement in capsule form leading to a significant increase in plasma antioxidant activity compared with when tea polyphenols were consumed as black tea or green tea (Henning et al., 2004). Second, an interesting finding noted that the addition of ascorbic acid (vitamin C) at an amount of 30mg to a 250mL tea beverage increased the catechin digestive recovery of epicatechin (EC), epigallocatechin (EGC), EGCG, and epicatechin-gallate (ECG) when tested using an in vitro digestion simulating gastric and small intestinal conditions. While it is uncertain whether or not this in vitro test will directly mimic in vivo ingestion of EGCG, these findings are partly the rationale for the inclusion of vitamin C to the Fat Smack™ ingredient profile. Finally, the use of green tea extracts has been reviewed for safety by the US Pharmacopeia (USP) Dietary Supplement Information Expert Committee (DSI EC) and this group concluded in their report that “Based on this safety review, the DSI EC determined that when dietary supplement products containing green tea extracts are used and formulated appropriately the Committee is unaware of significant safety issues that would prohibit monograph development” (Sarma et al., 2008). Moreover, a study using escalating dosages of EGCG indicated no adverse effects were noted at dosing up to 1600mg (Ullmann et al., 2003).

Cissus Quadrangularis
Cissus quadrangularis (CQ) is one of the most commonly used medicinal plants in Thailand and used in traditional African and Ayurvedic medicine. As with green tea extract, CQ renders multiple physiological effects, including potent antioxidant effects to minimize oxidative stress, anti-arthritis benefits, as well as anti-obesity properties. Specifically, the chemical constituents of CQ (e.g., flavonoids, phytosterols and keto-steroids) have shown promise as powerful antioxidants and demonstrated efficiency for lipase and amylase inhibition—collectively contributing to reduced oxidative stress and weight loss. At least three human studies have been conducted using this agent, all reporting beneficial effects on weight loss and associated variables. As an aside, although noted using an animal model, CQ stem extract was recently reported to provide hepatoprotection by exhibiting antioxidant and insulin-sensitizing activities (Chidambaram and Venkatraman, 2010). Coupled with the human evidence provided below, CQ appears to have promise as a health-enhancing, weight/fat loss agent.

In the first study using CQ (Oben et al., 2006), 123 overweight and obese individuals (47.2% male; 52.8% female; ages 19-50) were enrolled. The 92 obese subjects were randomized into three groups; placebo, CQ formulation/no diet, and CQ formulation/diet (2100-2200 calories/day). The 31 overweight subjects formed a fourth (no diet) treatment group. All subjects received two daily doses of the CQ formulation (CQ extract, soy albumin extract, green tea extract, niacin bound chromium, selenium, vitamin B6, vitamin B12, and folic acid) or placebo and remained on a normal or calorie-controlled diet for 8 weeks. Results indicated that statistically significant net reductions in body weight and central obesity, as well as in fasting blood glucose, total cholesterol, LDL-cholesterol, triglycerides, and C-reactive protein (a marker of systemic inflammation) were observed in subjects who received the CQ formulation, regardless of diet. HDL-cholesterol was also increased significantly. Collectively, these data indicate that CQ (in particular when combined with other ingredients as used in the present study) might be a viable candidate in a weight/fat loss product, as multiple health related effects are observed with treatment.

In a follow-up study conducted using CQ (Oben et al., 2007), a proprietary extract of CQ was compared to a proprietary formulation containing CQ on measures of weight, blood lipids, and oxidative stress (free radical mediated oxidation of large and small molecules) in a sample of overweight and obese subjects. Part one of the study investigated the in vitro antioxidant properties of the two conditions, while part two of the study included a double-blind placebo controlled design, enrolling initially 168 overweight and obese subjects (38.7% males; 61.3% females; ages 19-54), of whom 153 completed the study. Subjects received two daily doses of either condition or a placebo and were instructed to maintain their normal levels of physical activity. Results indicated that both conditions containing CQ exhibited antioxidant properties in vitro, as well as in vivo, as they resulted in significant reductions in plasma thiobarbituric acid reactive substances (TBARS) and protein carbonyls. Both conditions also brought about significant reductions in weight, body fat, total cholesterol, LDL-cholesterol, triglycerides, and fasting blood glucose levels over the intervention period, alongside an increase in both HDL-cholesterol and plasma 5-HT. Taken together, these findings indicate both weight/fat loss and cardio-protective properties of CQ supplementation.

The third study conducted using CQ involved an evaluation of the effects of two formulations, CQ-only and a CQ/Irvingia gabonensis combination, on weight loss in overweight and obese human subjects (Oben et al., 2008). The study involved a 10 week intervention period in which subjects (72 obese or overweight men and women (45.8% male; 54.2% female; ages 21-44; mean age = 29.3) were randomized using a double-blind, placebo-controlled design to three equal (n = 24) groups: placebo, CQ-only and CQ/Irvingia gabonensis combination. No major dietary or exercise changes were suggested during the study. Body weight, body fat (using bio-electrical impedance), waist size, total plasma cholesterol, LDL cholesterol, fasting blood glucose level were taken at baseline and at 4, 8 and 10 weeks. Results indicated that compared to the placebo group, the two active groups demonstrated a statistically significant difference on all six variables by week 10, with a greater change noted at week 10 compared to week 4 (even greater change was noted in the CQ/Irvingia gabonensis combination group). The decrease in body weight with CQ alone was approximately 19 pounds (~9%) in the 10 week intervention. While this is impressive, the most striking changes were noted in the reduction in blood cholesterol and glucose values—which admittedly, are so impressive they are almost difficult to believe (in particular for the CQ/Irvingia gabonensis combination).

Although subjects in this study were overweight or obese (BMI >25kg∙m-2), many individuals planning to use a weight/fat loss agent also fall into these categories; hence, these data are highly specific to many considering using Fat Smack™. These results agree with prior work in human subjects using CQ and suggest this ingredient may prove beneficial in overall weight loss over time, while providing cardio-protective properties specifically related to a reduction in oxidative stress and an improvement in blood lipids.

Considering the work performed by Oben and colleagues, the results for use of CQ are very impressive, especially in terms of the changes in bloodborne measures. A positive attribute of the Oben work is the fact that all studies included a relatively large sample size, something that is lacking in many dietary supplement intervention studies. Of course, the greater sample size also increases the likelihood of obtaining statistically significant findings (due to the increase in statistical power). Further studies, by different groups of investigators, are needed to corroborate the findings of Oben and coworkers. Lastly, few adverse events were noted in the above studies using CQ (e.g., headache, lack of sleep, gas), which were similar to those reported with use of placebo.

Capsaicinoids
Capsaicinoids are the major pungent, naturally occurring active compounds in capsicum fruits such as hot chili peppers—a fruit of the genus capsicum plant family. The most abundant forms of capsaicinoids found within hot red peppers are capsaicin (8-methyl-N-vanillyl-6-nonenamide), dihydrocapsaicin, and nordihydrocapsaicin (Thomas et al., 1998). If consuming whole hot peppers, approximately 3mg of capsaicinoids are present within 1g of dried red pepper (Yoshioka et al., 1995).

Capsaicinoid ingestion is typically associated with intense feelings of heat. In fact, the amount of heat generated by these capsaicinoids is typically measured using Scoville Heat Units (SHU), with pure capsaicin yielding the highest measure of 15,000,000 SCU. With such a high rating, very little capsaicin in needed to provide a measureable effect, and too much may be associated with gastrointestinal upset.

In terms of effects related to weight/fat loss, capsaicinoids have been reported in multiple studies to reduce ad libitum food intake, increase thermogenesis and energy expenditure, and enhance lipolysis (for a review please see Bloomer et al., 2009b). The lipolytic effect has been recently demonstrated using a very low dose capsaicinoid (Bloomer et al., 2010). Although the exact mechanisms are not yet fully understood, it is believed that capsaicinoids limit weight gain or may induce a weight loss via inhibiting adipogenesis, a finding noted using a cell culture study (Hsu and Yen, 2007). Capsaicinoids are thought to function to stimulate thermogenesis by activating the sarcoplasmic reticulum Ca+-ATP-ase (SERCA). Thermogenesis, as indicated earlier, refers to process of heat generation via the hydrolysis of ATP via SERCA. In the presence of capsaicinoids, SERCA becomes uncoupled (i.e., ATP hydrolysis occurs without the transport of calcium) leading to greater heat energy production (Mahmmoud, 2008). In terms of lipolysis, capsaicinoids are an agonist of the transient receptor potential vanilloid subfamily member 1 (TRPV1). TRPV1, via a number of steps, ultimately leads to an increased activation of the sympathetic nervous system leading to the release of epinephrine and norepinephrine from adrenal glands. These catecholamine hormones interact with hormone sensitive lipase, an enzyme involved in lipolysis and the mobilization of fatty acids from storage sites. The increase in catecholamine secretion may be responsible for the reduction in ad libitum food intake noted with capsaicinoid intake—which may be due to the anorectic effects of catecholamines (Russek et al., 1987).

It is important to note, as with data pertaining to other ingredients presented within this paper, that these findings have been noted in human subjects, consuming capsaicinoids (or red peppers; and sometimes CH-19 sweet peppers or non-pungent capsinoids) in oral form. As is the case for most ingredients, while not all studies have reported positive outcomes for the above variables, the majority of published work has noted positive effects related to at least one of the above outcome measures. Unfortunately, some manuscripts fail to report on the specifics of the ingested treatment (e.g., capsaicin or dihydrocapsaicin capsules, CH-19 sweet peppers, capsiate or dihydrocapsiate capsules, etc.) and rather state either pepper intake or capsule intake. Therefore, the exact dosage of capsaicinoids used is sometimes unknown. Despite this, epidemiological data appear to support an association between the consumption of capsaicinoid containing foods and a lower incidence of obesity (Wahlqvist and Wattanapenpaiboon, 2001). This finding, coupled with the specific effects noted above, qualify capsaicinoids as an integral ingredient for inclusion within Fat Smack™.

From a safety perspective, the lethal dose of capsaicinoids is estimated at 47.2mg/kg (mouse) (see www.sciencelab.com for Material Safety Data Sheet), an amount that is significantly higher than that used in any clinical study to date. Despite this fact, it should be noted that certain individuals may experience gastric discomfort and a burning sensation with intake and may view this as an adverse reaction. While it is sometimes noted that capsaicinoids promote gastric ulcers, this is refuted in a review on the topic (Satyanarayan, 2006) with the author noting that “investigations carried out in recent years have revealed that chilli or its active principle capsaicin is not the cause for ulcer formation but a benefactor.” Nonetheless, individuals with aversions to hot and spicy foods may not tolerate capsaicinoid intake well. Additional safety information can be found in the following article (Anonymous, 1998).

Piperine
Piperine is an extract of black pepper and appears to function in a similar manner as capsaicinoids (by binding the TRPV1 receptors). Piperine is often used within nutritional supplements as an aid in absorption (at a dosage of 5mg—the same dosage as used in Fat Smack™), as it may promote rapid absorption of certain nutrients (e.g., curcumin, vitamins, amino acids) from the gastrointestinal tract. Specifically, it has been used in combination with caffeine and capsaicinoids in a recent investigation in which the authors reported an increase in energy expenditure with supplement use (Ryan et al., 2009). Beyond this, in vitro studies have demonstrated effects ranging from enhanced antioxidant activity to protecting against oxidative damage via quenching free radicals (Srinivasan, 2007). For these purposes, piperine (which is a Generally Recognized as Safe [GRAS] ingredient) is included within Fat Smack™.

Vitamin/Mineral Mix
Fat Smack™ contains a vitamin/mineral matrix to enhance the antioxidant activity of the other ingredients (e.g., EGCG and CQ), as well as for the support of healthy thyroid function. Several of the ingredients provided within this matrix are known to support healthy thyroid (e.g., vitamin A, selenium, iodine, iron, and zinc) which is important for the regulation of metabolic rate (energy expenditure). It should be noted that while these nutrients are well-known to support healthy thyroid function, no outlandish claims are made in support of this matrix beyond this. That is, unlike some companies, PURUS LABS® is in no way making claims that this particular blend of thyroid supporting ingredients will elevate thyroid to “drug-induced” levels. This is unnecessary and potentially undesirable in a product such as Fat Smack™, which is strong enough considering the ingredient profile aside from the vitamin/mineral mix.

While certain agents are used within some nutritional supplements in an attempt to boost thyroid activity, and may be capable of doing so when studied in animal models at high dosages, little or no direct evidence is available to support their use in human subjects. Consider for example, 3,5-diiodo-L-thyronine, which has indeed been reported to have thyromimetic effects in vivo, but is routinely delivered to hypothyroid animals (often via IV injection) at doses from 2.5 to 10 µg/100g body weight (Moreno et al., 1998), an amount equivalent to multiple milligrams quantities in man. While this may not seem like a high dosage, the sheer price of this material renders this ingredient impossible to include at an efficacious amount; some supplement companies choose to use small MICROgram amounts of this ingredient within their proprietary blends. While it is possible that a much lower dosage could provide some beneficial effect, this has yet to be supported by clinical study.

Besides the above, there is no evidence related to the long-term impact of many such agents on thyroid function. It is possible, as with many other exogenous agents administered in an attempt to boost natural hormone production, that the body may down-regulate endogenous production of thyroid hormone over time if chronically supplementing with a supposed thyroid hormone booster. Due to these issues, PURUS LABS® has simply decided not to focus specifically on attempting to boost thyroid function by using “novel,” supposed thyroid stimulators; instead, Fat Smack™ is fueled by a combination of ingredients thought to support healthy thyroid function in human subjects. Perhaps data pertaining to the use of naturally occurring agents that stimulate thyroid function in human subjects will become available in coming years, at which time they will be systematically investigated for potential inclusion within a revised Fat Smack™. However, at this time, the data simply are not available.

SUMMARY
Fat Smack™ contains a carefully selected blend of ingredients, based on human evidence for effect in relation to multiple aspects of weight/fat loss, and used at the research-proven dosages. While regular exercise and optimal dietary intake inclusive of frequent, macronutrient balanced and nutrient dense meals (as well as adequate water intake) should be viewed as most important in the quest for body weight/fat reduction and the reduction of cardiovascular and metabolic disease risk, use of a weight/fat loss agent such as Fat Smack™ may be an adjunct to this lifestyle plan.

As with all nutritional supplements, potential users should only use Fat Smack™ under the guidance of their personal physician. Individuals considering using Fat Smack™ should be healthy and over the age of 21. They should not have any diagnosed medical conditions or family history of medical conditions related to, but not limited to, high blood pressure, heart disease, stroke, or any other disease of cardiovascular or metabolic origin. Due to the pepper extracts used in this product, individuals with aversions to spicy foods may need to limit use of the product. Individuals subjected to anti-doping regulation by their athletic organizations should be advised that the 2-amino-4-methylhexane used within this product (and several other products currently sold on the supplement market) is currently considered a banned substance by certain athletic organizations. Individuals should review the product nutrition panel and label for information regarding the ingredients, dosing, and precautions for use. For more information on PURUS LABS® and its other performance/physique-enhancing products, please visit www.puruslabs.net.

Overview and Rationale for use
Before we begin the discussion of PURUS LABS™ SLINshot™ it is important to understand that while other ingredients targeting glucose regulation appear promising in their own right, PURUS LABS™ has made the careful decision to use Artemisia Dracunculus L. var. inodora exclusively within SLINshot™. The reason for this decision is threefold. First, Artemisia Dracunculus L. var. inodora has been used in several studies to date with results that all essentially parallel each other; that is, the studies support one another rather than refute each other. This is often not the case when studying nutritional ingredients. Second, when designing a dietary supplement PURUS LABS™ believes that not only do the appropriate ingredients need to be included within the product, but the correct and efficacious dosage of ingredient needs to be included. Each single serving of SLINshot™ contains 1000mg of Artemisia Dracunculus L. var. inodora. If several ingredients were to be included within SLINshot™ simply to compete with other companies attempting to fill the label with ingredients, the dosage of each ingredient would need to be significantly lower than the believed effective dosage. This makes little scientific sense and is not something PURUS LABS™ will ever do. Alternatively, if all (or many) of the ingredients that appear to have some promise in terms of glucose regulation were included at the believed effective dosage, consumers would need to consume 10-12 capsules per serving rather than 2 capsules, and the per unit cost of the product would be 5-6 times higher. Again, when considering the potential benefit of doing this based on the currently available evidence for effect, it simply does not make sense. PURUS LABS™ believes the exclusive use of Artemisia Dracunculus L. var. inodora within SLINshot™ will provide consumers with a scientifically sound, effective product at a reasonable intake level and cost. This dietary supplement may serve the dual purpose to allow for enhanced glucose and creatine uptake into tissue. Third, PURUS LABS™ has personally experimented with Artemisia Dracunculus L. var. inodora (aqueous extract) and has observed the effects of this ingredient first hand. Of course, further well-controlled research is necessary to provide confirmation for the proposed effects in human subjects.

Preliminary Human Subject Findings
As mentioned earlier in the paper, the majority of work involving Artemisia Dracunculus L. var. inodora has been conducted in animals. While findings from animals can often be interesting and sometimes translate to humans, this is not always the case. This is especially true considering the route of administration in many animal studies is not oral (like that for human dietary supplements), and the dosage of ingredient used in many animal studies is often far greater than most humans will want to consume or could afford to use. Fortunately, Artemisia Dracunculus L. var. inodora has been delivered to animals via oral ingestion and at a dosage that is actually manageable in terms of translation to human daily ingested amounts. Moreover, Artemisia Dracunculus L. var. inodora has been previously used by humans at an oral dosage of 1000mg, with favorable effects noted for creatine clearance from plasma (Jäger et al., 2008).

Considering the work of Jäger and colleagues (2008) using Artemisia Dracunculus L. var. inodora to enhance creatine clearance from plasma, the effects of both 1000mg (equivalent to 1 serving of SLINshot™) and 2000mg (equivalent to 2 servings of SLINshot™) of Artemisia Dracunculus L. var. inodora on blood glucose following an OGTT has been studied using a case analysis in an effort to generate pilot data to guide future research studies (unpublished data). The text below describes this work.

The Experiments Described
For data collected from experiments described here please refer to Table 1 and Figure 2. Two adult resistance trained men (Subject X: 225 pounds, 10% body fat; Subject Y: 180 pounds, 7% body fat) performed two OGTTs with and without experimental research grade SLINshot™. Subject X used 1000mg (1 serving) of SLINshot™ and subject Y used 2000mg (2 servings) of SLINshot™. The SLINshot™ was ingested 15 minutes prior to consuming 75 grams of dextrose solution (300 calories). This is standard procedure for a clinically administered OGTT. Blood samples were collected from subjects before ingesting the SLINshot™ (after a 10 minute quiet rest period) and at 15, 30, 45, and 60 minutes following ingestion of the dextrose solution. (Note: As opposed to a blood collection every 30 minutes and a duration of collection of 2-3 hours, often employed for an OGTT administered in a clinical setting, it was decided to obtain more frequent blood samples but to cease measurements at one hour post ingestion. This decision was based on prior work involving young, healthy subjects performing an OGTT—for which blood glucose more quickly returns to pre-meal values as compared to individuals with impaired glucose tolerance). During the 60 minute post ingestion period subjects remained relaxed and ingested no additional food or calorie containing beverages. Following blood sample collection, glucose was analyzed in serum using standard enzymatic procedures. Assays were performed in triplicate.

Table 1 provides values for each time point of sample collection for both placebo and SLINshot™ trials. The area under the curve (AUC) is also presented which uses a mathematical model to represent the “sum” over the course of the one hour post ingestion period. Percent difference values between placebo and SLINshot™ trials are included for each time point as well as for AUC. Figure 2 provides a graphical representation of the data. As can be seen, oral intake of SLINshot™ provides for blood glucose disposal effects in healthy men averaging 20.5%.

Pertaining to the above, while only two subjects were used in this case study analysis, it cannot be concluded that all individuals will respond in the same manner. Clearly, additional research using a sample of 10-20 individuals is needed to provide further support for these initial findings. This is true for all herbal forms of glucose regulatory agents recommended for use in human subjects (Cefalu et al., 2008b). However, when collectively considering the evidence from both animals and humans (in relation to both glucose and creatine clearance), the results are indeed noteworthy.

It should be understood, as with all dietary supplements, individual results to treatment may vary. Therefore, individuals may require more or less than 1 serving of SLINshot™ in order to achieve the desired result (i.e., glucose or creatine clearance). Keep in mind that in much of the work related to Artemisia Dracunculus L. var. inodora, test subjects/animals consumed 75 grams of research grade dextrose (a simple sugar), or the equivalent. Individuals not consuming such high amounts of carbohydrate (in particular simple sugar) may not require as much SLINshot™ in order to optimally manage blood glucose levels (or may not require any glucose disposal agent at all). However, SLINshot™ was developed with the idea that athletes would use this supplement along with their post-exercise carbohydrate meal which typically contains a significant amount of carbohydrate (often in the form of simple sugars). In this situation, it is recommended that SLINshot™ be used at 1000-2000mg (1-2 servings) depending on age, glucose tolerance, and overall health status. For example, subjects X and Y experienced similar effects of SLINshot™ despite two different dosages being used. As with all dietary supplements, experimentation by each individual is suggested. This applies to both athletes and non-athletes and should be considered relative to meals associated with exercise bouts as well as those outside of the context of acute exercise.

Conclusions and Practical Applications
Management of blood glucose following feeding is of importance for athletes and non-athletes. In relation to the former group, it is possible that post exercise feedings that contain high amounts of carbohydrate for purposes of glycogen replenishment may better be taken up into tissue via dietary support in the form of SLINshot™ (Artemisia Dracunculus L. var. inodora). Moreover, this ingredient may facilitate uptake of other nutrients commonly used by athletes (e.g., creatine). Collectively, the product SLINshot™ from PURUS LABS™ may be used as an adjunct to an already well-designed exercise and nutrition plan targeting optimal blood glucose regulation and/or glycogen supercompensation. Daily use of this product in conjunction with high carbohydrate meals, either post workout or at other times of the day as needed, may aid in heightened glucose clearance from the blood and uptake into tissue. This may not only improve glycogen replenishment in athletes seeking this outcome but may also maintain overall health as related to optimal blood glucose management (e.g., reduce the potential harmful effects of free radicals, improve energy and mood, reduce excess body fat accumulation, etc).

As with all nutritional supplements, potential users should consult their personal physician prior to using SLINshot™. This especially applies to those individuals with known problems related to blood glucose regulation. In addition, potential users should review the product nutrition panel and label for information regarding the ingredient, dosing, and precautions for use. For more information on PURUS LABS™ and its other performance/physique-enhancing products, please visit www.puruslabs.net.

Zuberi AR. Strategies for assessment of botanical action on metabolic syndrome in the mouse and evidence for a genotype-specific effect of Russian tarragon in the regulation of insulin sensitivity. Metabolism. 2008 Jul;57(7 Suppl 1):S10-5.

D-pol™ Overview and Rationale for use
d-pol™ is a new product designed by PURUS LABS®, which adheres to the established guidelines of PURUS LABS® in terms of including only those ingredients that have been used in humansubjects via oralingestion, and reported to promote favorable outcomes in relation to the target areas of interest (e.g., enhanced exercise performance, increased circulating testosterone—potentially leading to increased lean body mass, enhanced overall health). d-pol™ contains a unique blend of ingredients, each carefully selected for their potential impact on physical performance and human health—included at the precise dosage necessary to promote the desired effect (as supported by peer reviewed scientific evidence obtained from studies using human subjects). The text below outlines the ingredients used within d-pol™, while providing a brief summary of the rationale for use.

d-aspartic acid
Elevating circulating testosterone has been, is now, and will likely continue to be the objective of many hard training athletes. It is well known that testosterone is associated with gains in both muscle size and strength, in addition to having positive effects on several other vital components of human health. One dietary ingredient reported to produce measureable gains in circulating testosterone is d-aspartic acid (DAA). d-aspartic acid has been reported in multiple animal and in vitro studies to be involved in testosterone synthesis, in species ranging from boar (Lamanna et al., 2007a) to lizard (Raucci et al., 2005). It also stimulates the production of other hormones such as prolactin, leutinizing hormone (LH), and growth hormone. Finally, DAA acts as a neurotransmitter/neuromodulator (D'Aniello, 2007)—an effect noted in several studies using animal models and in vitro experiments.

Considering the sound evidence for a role of DAA in testosterone synthesis, a human study was recently performed to assess the effectiveness of oral DAA supplementation with regards to testosterone elevation (Topo et al., 2009). Subjects included 43 men (27-37 yrs), 20 who received a placebo and 23 who received a DAA supplement (3.12 grams of DAA combined with B-vitamins—the same dosage of DAA provided within d-pol™). The supplement was delivered orally for 12 days. Following the 12 days of treatment with the supplement, 20 of the 23 subjects (87%) had significantly higher circulating LH levels, with an average increase of 33%. With regards to circulating testosterone, 20 of the 23 subjects (87%) had significantly higher values at day 12 compared to pretreatment, with an average increase of 42%.
For both LH and testosterone, the changes were time dependent—a greater increase was observed on day 12 compared to day 6 of treatment. Therefore, it is possible that continued treatment with the supplement may have resulted in a further increase in these variables. Although measurements of muscle size and strength were not included in this study, it is possible that the increase in circulating testosterone may be associated with a measureable increase in these variables. Further work is needed to confirm these effects.

While the above findings are interesting and may prove beneficial to those seeking a natural method of increasing circulating testosterone, it should be noted that DAA has been reported to increase aromatase activity, as measured using boar testes (Lamanna et al., 2007b). At the present time it is unknown what amount of DAA would need to be orally ingested by humans in order to promote this effect. No adverse outcomes were noted in the Topo et al. (2009) investigation. Regardless, as with other testosterone stimulating agents, a cyclic schedule of supplementation of d-pol™ is recommended. If adhering to this recommendation, and when considering the inclusion of vitamin D within d-pol™, it may be wise to cycle “off” during periods of optimal sunlight exposure (when natural vitamin D synthesis would be highest—see text below for additional information pertaining to this).

B-vitamins
B-vitamins constitute an entire group of water soluble vitamins, with multiple known
biochemical effects within the body—as evidenced by the thousands of scientific reports
documenting such effects. B-vitamins have known effects in relation to increasing the rate of
metabolism, enhancing immune function, and promoting cell growth and division. Of greatest
importance in relation to the use of these vitamins within d-pol™, the B-vitamins have been used
in combination with DAA for purposes of elevating circulating testosterone: B9 (folic acid),
B6 (pyridoxine), and B12 (cyanocobalamin). Because dietary folic acid is not well metabolized to
the active form by some individuals due to a genetic defect within the folate pathway, the
naturally occurring form of this vitamin (folate) is included within d-pol™.

Nitrate
Nitrate is an inorganic anion abundant in vegetables (e.g., beets, spinach) and converted within the body to nitrite in the entero-salivary circulation (Duncan et al., 1995). The study of nitrate, nitrite, and bioactive nitrogen oxides including nitric oxide, is of great interest to investigators, due to the multiple biological roles of the nitrate-nitrite-nitric oxide pathway (Lundberg et al., 2009).

A relatively new area of investigation is now focused on the use of nitrate supplementation for purposes of improving exercise performance (Bailey et al., 2009; 2010; Lansley et al., 2010; Larsen et al., 2007; 2010; Vanhatalo et al., 2010). In general, the findings from this work indicate an increase in circulating levels of nitrite following nitrate supplementation (possibly suggesting an increase in circulating nitric oxide), in addition to a lower oxygen cost during exercise. Interestingly, these findings are observed after just a few (e.g., 4-7) days of nitrate supplementation. Such results may translate into lower perceived effort at any given submaximal workload or an increase in the actual amount of work that subjects are able to perform during a given exercise session.

While this area of research remains in its infancy, additional studies inclusive of outcome measures beyond blood nitrite and oxygen cost of exercise are needed to more fully detail the ergogenic benefits of nitrate supplementation. To date, the majority of this work has been performed in the laboratory of Professor Andrew Jones at Exeter University in the UK, using beetroot juice as the delivery vehicle for nitrate. In much of this work, subjects ingest 500mL of beetroot juice (providing approximately 350mg of dietary nitrate—the same dosage provided within d-pol™). Other work has used dietary nitrate directly (e.g., sodium nitrate), with the dosage based on subjects’ body mass.

As stated, findings from these studies indicate an increase in circulating levels of nitrite and a reduction in the oxygen cost during exercise. While most exercise studies have involved aerobic work, one recent investigation has focused on resistance exercise (Bailey et al., 2010). In this study, subjects received beetroot juice for six consecutive days and completed a series of low-intensity and high-intensity "step" exercise tests on the last 3 days of supplementation for the determination of the muscle metabolic (using (31)P-MRS) and pulmonary oxygen uptake (VO2) responses to exercise. As is typical, on days 4-6, beetroot juice resulted in a significant increase in blood nitrite. During low-intensity exercise, beetroot juice attenuated the reduction in muscle phosphocreatine concentration and the increase in VO2. During high-intensity exercise, beetroot juice reduced VO2 slow components and significantly improved exercise time to exhaustion. Moreover, the total rate of ATP turnover was estimated to be less for both low-intensity and high-intensity exercise. The authors concluded that the reduced oxygen cost of exercise following nitrate supplementation appears to be due to a reduction in the ATP cost of muscle force production. These findings are further supported by a recent investigation noting an improvement in mitochondrial efficiency with nitrate supplementation (Larsen et al., 2011), a finding that is also correlated to the reduction in oxygen cost during exercise. Collectively, these changes appear to allow exercise to be tolerated for a greater period of time—which may translate into more repetitions being performed during a given set of exercise and/or a greater load to be used during each set of exercise. For aerobic exercise bouts, the duration of exercise may be increased following supplementation. These are indeed findings of interest to any exercise enthusiast.

Aside from a performance benefit, nitrate supplementation has been reported to lower blood pressure in humans (Kapil et al., 2010; Vanhatalo et al., 2010; Webb et al., 2008), as discussed recently (Ferreira & Behnke, 2010; Gilchrist et al., 2011). Dietary nitrate has been reported to prevent endothelial dysfunction induced by an acute ischemic insult in the human forearm and to attenuate ex vivo platelet aggregation in response to collagen and ADP (Webb et al., 2008). These effects, coupled with the vasodilatory effects of increased circulating nitrite, seem to be responsible for the blood pressure lowering effect of dietary nitrate. While this may not be of primary importance to many athletes, a reduction in blood pressure should at least be of interest to individuals, in particular those who use bodybuilding drugs known to cause elevations in both resting and stress-induced blood pressure. From a general health point of view, a slight reduction in blood pressure is correlated to a lower risk of cardiovascular disease.

In addition to the above benefits of nitrate supplementation, it has been noted that nitrate may also improve the absorption of certain nutrients. While this appears of some interest, because such findings are isolated to designs not involving oral ingestion of nitrate by human subjects, the potential for improved absorption of nutrients following nitrate ingestion it is not a major consideration for the inclusion of nitrate within d-pol™. If such effects on enhanced nutrient absorption are apparent with nitrate ingestion, this would simply be an adjunct to the well-documented performance and health benefits that are observed.

Finally, it is important to address the potential safety concern over ingestion of dietary nitrate, which has been raised (Derave & Taes, 2009; Petróczi & Naughton, 2010), with more specific concern over ingestion of high amounts of dietary nitrite (Lundberg et al., 2011). This has been appeased in recent years by experts working specifically in this area of research (Benjamin et al., 2009; Gilchrist et al., 2010). The general consensus is that ingestion of low amounts of dietary nitrate appears safe and may be associated with improvements in parameters of health (e.g., vascular function) and physical performance.

Vitamin D
Vitamin D is a fat-soluble vitamin with multiple known functions within the body (Holick, 2004; Sutton & MacDonald, 2003; Thacher & Clarke, 2011), ranging from the maintenance of normal calcium metabolism to a modulator of cellular immunity. Vitamin D3 (cholecalciferol) can be synthesized by humans in the skin upon exposure to ultraviolet-B radiation from sunlight. Vitamin D can also be obtained from the diet. Unfortunately, vitamin D is found naturally in very few foods. According to the Linus Pauling Institute, “foods containing vitamin D include some fatty fish (mackerel, salmon, sardines), fish liver oils, and eggs from hens that have been fed vitamin D. In the U.S., milk and infant formula are fortified with vitamin D so that they contain 400 IU (10 mcg) per quart. However, other dairy products, such as cheese and yogurt, are not always fortified with vitamin D. Some cereals and breads are also fortified with vitamin D. Recently, orange juice fortified with vitamin D has been made available in the U.S.”

Based on the above, vitamin D supplementation is likely required by most individuals. This is particular true if exposure to natural sunlight is limited and/or the use of sunscreen in routine. In fact, it has been reported that the majority (87%) of adults living in the relatively sunny climate of Tennessee are deficient in vitamin D (Long et al., 2011); a finding that is well supported by numerous investigations including adults (Holick, 2007), in addition to those including children. Individuals residing in parts of the country where the amount and intensity of sunlight is less than average may be at even greater risk for deficiency. Without adequate dietary intake and/or natural synthesis of vitamin D, multiple components of physical health may be compromised—again, a well documented outcome that has fueled the continued growth of vitamin D research.

Activated vitamin D (calcitriol) has been referred to as a pluripotent pleiotropic secosteroid hormone (Cannell et al., 2009). As a steroid hormone regulating more than 1000 vitamin D-responsive human genes, vitamin D may influence athletic performance. Most of the studies supporting the role of vitamin D with regards to exercise performance involved older adults. However, as noted by Cannell and colleagues (2009) in a recent review article on the topic “physical and athletic performance is seasonal; it peaks when 25-hydroxy-vitamin D levels peak, declines as they decline, and reaches its nadir when 25-hydroxy-vitamin D levels are at their lowest.” These findings are underscored by the potential impact that vitamin D has on skeletal muscle, as described recently (Bartoszewska et al., 2010; Ceglia, 2009; Hamilton, 2010). Furthermore, a recent study (Pilz et al., 2011) in healthy men who supplemented with 3332 IU of vitamin D daily for one year, noted a significant increase in total, bioactive, and free testosterone levels. Although this finding requires confirmation through further research, it appears as though some of the noted health-related effects of vitamin D supplementation in men may be mediated through an increase in testosterone. Considering the well-described health effects of vitamin D, coupled with the potential impact of vitamin D on exercise performance, supplementation with this powerful vitamin should be considered by all individuals.

Research indicates that vitamin D toxicity is very unlikely in healthy individuals when used at amounts less than 10,000 IU/day (Vieth, 1999). However, the Food and Nutrition Board of the Institute of Medicine (IOM) and a recent Clinical Practice Guideline from the IOM in conjunction with the Endocrine Society (Holick et al., 2011) has set a conservative tolerable upper intake level of 4,000 IU/day (100 mcg/day) for all adults. This is similar to the dosage used in many clinical trials and is the dosage provided within d-pol™.

Cofactors for Vitamin D
For optimal vitamin D absorption, taking vitamin D with the largest meal of the day seems most appropriate (Mulligan & Licata, 2010). Beyond this, it has been suggested that certain vitamin and mineral cofactors are potentially needed for optimal vitamin D action and effect. However, it should be noted that the need for these cofactors is debatable. Moreover, most of these co-factors are present in vitamin/mineral supplements, which most individuals consume regularly. The cofactors of interest include magnesium, zinc, vitamin K2, vitamin A, and boron. With regards to boron, aside from acting as a cofactor for vitamin D, one recent report indicates a testosterone elevating effect of this mineral (Naghii et al., 2010). It is possible that this may enhance the action of DAA on circulating testosterone. However, an earlier report for boron refutes these findings (Green and Ferrando, 1994). Collectively considering the above, boron and other co-factors are not included within d-pol™.

SUMMARY
d-pol™ contains a carefully selected blend of ingredients, based on human evidence for effect in relation to multiple aspects of health and performance. While regular and strenuous exercise, as well as optimal dietary intake inclusive of frequent, macronutrient balanced and nutrient dense meals (as well as adequate water intake) should be viewed as most important in the quest for optimal health and physical functioning, use of a dietary supplement such as d-pol™ may be an adjunct to this lifestyle plan. The use of d-pol™ should be considered in a single dosage with the largest meal of the day.

As with all nutritional supplements, potential users should only use d-pol™ under the guidance of their personal physician. Moreover, because d-aspartic acid (a chief ingredient in d-pol™) has been reported to increase circulating testosterone, d-pol™ should not be used by women. Individuals considering using d-pol™ should be healthy and over the age of 21. Users of d-pol™ should review the product nutrition panel and label for information regarding the ingredients, dosing, and precautions for use. For more information on PURUS LABS® and their other performance/physique-enhancing products, please visit www.puruslabs.net.